Efficient preparation of c-di-AMP at gram-scale using an immobilized Vibrio cholerae dinucleotide cyclase DncV

Sun, Qichao; Lv, Yuzhen; Zhang, Chenhui; Wu, Weifang; Zhang, Rui; Zhu, Chunyuan; Li, Yaoyao; Yuan, Huiqing; Zhu, Jing; Zhu, Deyu

doi:10.1016/j.enzmictec.2020.109700

Cited by 5 publications

(4 citation statements)

References 30 publications

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“…The yield of natural CDN from chemical synthesis is low and the synthesis route is complex. At present, gram-grade c-di-AMP can be prepared in the laboratory using immobilized Vibrio cholerae dinucleotide cyclase DncV, and can be used on a large industrial scale in the future with expected total yields of nearly 80%, thus facilitating the enzymatic synthesis of CDNs in large quantities ( Sun et al, 2021 ; Bartsch et al, 2022 ). To obtain a variety of CDNs, different DncV homologs can be utilized for different purposes ( Novotna et al, 2021 ).…”

Section: Discussionmentioning

confidence: 99%

Cyclic dinucleotides mediate bacterial immunity by dinucleotide cyclase in Vibrio

Zhou

et al. 2022

Front. Microbiol.

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The cyclic GMP-AMP (cGAMP) synthase (cGAS) recognizes cytosolic DNA and synthesizes the second messenger, cGAMP, thus activating the adaptor protein stimulator of interferon genes (STING) and initiating the innate immune responses against microbial infections. cGAS-STING pathway has been crucially implicated in autoimmune diseases, cellular senescence, and cancer immunotherapy, while the cGAS-like receptors in bacteria can protect it against viral infections. Dinucleotide cyclase in Vibrio (DncV) is a dinucleotide cyclase originally identified in Vibrio cholerae. The synthesis of cyclic nucleotides by DncV, including c-di-GMP, c-di-AMP, and cGAMP mediates bacterial colonization, cell membrane formation, and virulence. DncV is a structural and functional homolog of the mammalian cytoplasmic DNA sensor, cGAS, implicating cGAS-STING signaling cascades may have originated in the bacterial immune system. Herein, we summarize the roles of DncV in bacterial immunity, which are expected to provide insights into the evolution of cGAS-STING signaling.

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Section: Discussionmentioning

confidence: 99%

Cyclic dinucleotides mediate bacterial immunity by dinucleotide cyclase in Vibrio

Zhou

et al. 2022

Front. Microbiol.

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“…The enzyme btDisA from Bacillus thuringiensis was used to synthesize 100 mg of c‐di‐AMP in 4 h in a 50 ml reaction system and hence proved the possibility for a gram‐scale production (Zheng et al, 2013). Recently, an immobilized form of the cyclic‐AMP‐GMP synthetase DncV from Vibrio cholerae was used for the production of c‐di‐AMP at a gram‐scale resulting in titers of up to 3.3 g L −1 and a process yield of 80% (Table 1) (Sun et al, 2020). The biocatalyst DncV was previously reported to catalyze the synthesis of the bacterial cGAMP containing two 3′5′‐linkages (Launer‐Felty & Strobel, 2018).…”

Section: Process Aspects Of Biocatalytic Cdn Productionmentioning

confidence: 99%

“…This sequential chromatography resulted in 42.8 mg purified 2′3′‐cGAMP from 0.5 L culture, 53% purification yield, and a purity of 99.5% (w/w) measured via UV spectroscopy. The highest overall yield of 80% was obtained by an integrated process development of c‐di‐AMP production (Sun et al, 2020). The process was performed with immobilized DncV that was easily separated after reaction.…”

Section: Process Aspects Of Biocatalytic Cdn Productionmentioning

confidence: 99%

Biotechnological production of cyclic dinucleotides—Challenges and opportunities

Bartsch

Becker

Rolf

et al. 2022

Biotech & Bioengineering

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Cyclic dinucleotides (CDNs) are widely used secondary signaling molecules in prokaryotic and eukaryotic cells. As strong agonists of the stimulator of interferon genes, they are of great interest for pharmaceutical applications. In particular, cyclic-GMP-AMP and related synthetic CDNs are promising candidates in preclinical work and even some in clinical phase 1 and 2 studies. The comparison of chemical and biocatalytic synthesis routes elucidated that biological CDN synthesis offers some advantages, such as shorter synthesis time, avoiding complex protective group chemistry, and the access to a new spectrum of CDNs. However, the synthesis of CDNs in preparative quantities is still a challenge, since the chemical synthesis of CDNs suffers from low yields and complex synthetic routes and the enzymatically catalyzed synthesis is limited by low product titers and process stability. We aim to review the latest discoveries and recent trends in chemical and biocatalytic synthesis of CDNs with a focus on the synthesis of a huge variety of CDN derivatives. We furthermore consider the most promising biotechnological processes for CDN production by evaluating key figures of the currently known processes.

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“…Owing to its signi cant functions, cAMP was extensively applied in treatment of cardio-cerebrovascular diseases and area of animal feed additive attracting extensively attentions (Liu et al 2018). cAMP can be produced via three different processes: chemical synthesis (Lelle et al 2019), enzymatic (Sun et al 2021), and fermentation processes (Niu et al 2018). Among these approaches, fermentation production by microorganism has attracted increasing attention, because the process is economical, sustainable and environmental-friendly as compared with complicated chemical synthesis.…”

Section: Introductionmentioning

confidence: 99%

Enhanced endogenous amino acids and energy metabolism level for cAMP biosynthesis by Arthrobacter sp. CCTCC 2013431 with citrate as cosubstrate

Chen

et al. 2021

Biotechnol Lett

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Objectives In our previous study, citrate was used as auxiliary energy substance for improving cAMP fermentation performance, however, the regulation mechanism of citrate on improved cAMP contents was not clear. To elucidate the regulation mechanism, cAMP fermentations with/without citrate addition were conducted in a 7 L fermentor using Arthrobacter sp. CCTCC 2013431 and assays on key enzymes activities, energy metabolism level, amino acids contents and peroxidation level were performed.Results With 3 g/L-broth sodium citrate added, cAMP concentration and conversion yield from glucose reached 4.34 g/L and 0.076 g/g which were improved by 30.7% and 29.8%, respectively, when compared with those of control. Citrate changed carbon ux distribution among different routes and more carbon ux was directed into pentose phosphate pathway bene cial to cAMP synthesis. Meanwhile, energy metabolism together with precursor amino acids levels were improved signi cantly owing to strengthened metabolic intensity of tricarboxylate cycle by exogenous citrate utilization which provided energy and substance basis for cAMP production. Moreover, higher glutamate synthesis and oxidative stress caused by citrate addition consumed excessive NADPH derived from pentose phosphate pathway by which feedback suppression for pentose phosphate pathway was relieved e ciently.Conclusion Citrate promoted cAMP fermentation production by Arthrobacter sp. CCTCC2013431 due to enhanced precursor amino acids, energy metabolism level and relieved feedback suppression for pentose phosphate pathway.

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Efficient preparation of c-di-AMP at gram-scale using an immobilized Vibrio cholerae dinucleotide cyclase DncV

Cited by 5 publications

References 30 publications

Cyclic dinucleotides mediate bacterial immunity by dinucleotide cyclase in Vibrio

Cyclic dinucleotides mediate bacterial immunity by dinucleotide cyclase in Vibrio

Biotechnological production of cyclic dinucleotides—Challenges and opportunities

Enhanced endogenous amino acids and energy metabolism level for cAMP biosynthesis by Arthrobacter sp. CCTCC 2013431 with citrate as cosubstrate

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